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Comparison of the Kato-Katz technique, hatching test and indirect hemagglutination assay (IHA) for the diagnosis of Schistosoma japonicum infection in China.

The Kato-Katz technique (duplicate 41.7 mg fecal smears), hatching test and indirect hemagglutination assay (IHA) were compared for their ability to detect human Schistosoma japonicum infection in two endemic villages (Zhonjiang and Zhuxi) in rural China. The hatching test (using a nylon bag, and based on about 30 g of feces) and IHA are conventional Chinese diagnostic methods. In both villages, the trends of prevalences with age and sex were comparable for the different methods. In Zhuxi, Kato-Katz examinations of stools from 7 different days and hatching were available, which could be used as a reliable gold standard. This resulted for IHA in a sensitivity of 80% and a specificity of 48%. The sensitivity of the Kato-Katz technique using one stool specimen was 68%, twice that of hatching (33%). In Zhonjiang, however, hatching resulted in more positive cases than Kato-Katz (prevalence 31% vs. 24%). Apparently, the result of the hatching test depends on environmental factors such as temperature and water quality. Although imperfect, Kato-Katz is recommended out of the three evaluated techniques as the method of choice for large-scale screening of S. japonicum. Hatching is much more tedious, provides inconsistent and only qualitative results, and is not much more sensitive than Kato-Katz. Its poor specificity makes IHA unsuitable for individual screening, but it may be more effective for community diagnosis.

Adolescent↗

Bacisubin, an antifungal protein with ribonuclease and hemagglutinating activities from Bacillus subtilis strain B-916.

An antifungal protein, with a molecular mass of 41.9 kDa, and designated as bacisubin, was isolated from a culture of Bacillus subtilis strain B-916. The isolation procedure consisted of ion exchange chromatography on DEAE-Sepharose Fast Flow, and fast protein liquid chromatography on Phenyl Sepharose 6 Fast Flow and hydroxyapatite columns. The protein was adsorbed on all three chromatographic media. Bacisubin exhibited inhibitory activity on mycelial growth in Magnaporthe grisease, Sclerotinia sclerotiorum, Rhizoctonia solani, Alternaria oleracea, A. brassicae and Botrytis cinerea. The IC50 values of its antifungal activity toward the last four fungal species were 4.01 microM, 0.087 microM, 0.055 microM and 2.74 microM, respectively. Bacisubin demonstrated neither protease activity, nor protease inhibitory activity. However, it manifested ribonuclease and hemagglutinating activities.

Amino Acid Sequence↗

Cross-reactivity of influenza A (H3N2) hemagglutination-inhibition antibodies induced by an inactivated influenza vaccine.

The antigenic drift of influenza A (H3N2) virus in 2003-2004 necessitated a change in the vaccine from the A/Panama to the A/Wyoming strain for the 2004-2005 season. Using hemagglutination inhibition, we therefore tested antibodies in sera of 39 individuals (mean age 64.6 years) at the end of the 2003-2004 season for cross-reactivity to vaccine strains and H3N2 antigens subject to antigenic drift. Antibodies against both A (H3N2) Panama and Wyoming developed in 5/13 (38.5%) unvaccinated individuals, whereas, 22/26 (84.6%) vaccinees developed antibodies to Panama and 21/26 (80.8%) to Wyoming. None of these individuals suffered an influenza episode that season. The results suggest that the elderly might develop protective levels of cross-reactive A (H3N2) Wyoming HI antibodies following vaccination with the Panama strain. Such strains, like the ones included in the 2003-2004 influenza vaccine, might be expected to provide a broad-spectrum antibody response that could be effective even in the face of single season antigenic drift.

Aged↗

Presence of hemagglutination inhibition and neutralization antibodies to Japanese encephalitis virus in wild pigs on an offshore island in Singapore.

A study was conducted to determine the presence of hemagglutination inhibition (HI) and neutralization antibodies against Japanese encephalitis virus (JEV) in wild pigs on an offshore island in Singapore. Blood samples were collected from 28 wild pigs on the island. All the sera tested with HI assay and plaque reduction neutralization tests were found to be positive for antibodies against JEV. Our results indicate that the wild pigs have been infected with JEV on the offshore island and there is JEV transmission.

Animals↗

Detection of anti-zona pellucida activities in human sera by the passive hemagglutination reaction.

A simple method of detecting anti-zona activities in human sera has been developed by passive hemagglutination reaction ( PHAR ) using bovine erythrocytes as indicator cells coated with purified porcine zona substance. By this test, 8 of 88 serum samples (9.1%) from infertile women gave positive reactions, whereas only 1 of 90 control sera (1.1%). A positive immunofluorescence (IF) assay confirmed the results of the PHAR in seven of the eight positive samples. Positive PHAR and IF on porcine zonae were abolished after adsorption with porcine erythrocytes, but IF on human zonae was retained by four of eight serum samples, indicating the presence of anti-zona autoantibodies in these infertile sera that were directed to an antigen(s) proper to human zonae.

Adult↗

A hemagglutination test for binding of hydrazide-derivatized cytochemical markers.

A sensitive hemagglutination test was developed in order to predict the ability of amine- or hydrazide-derivatized cytochemical markers to interact with cell surface aldehydes in various in vitro and in situ systems. Human red blood cells were modified by glutaraldehyde fixation and subsequent oxidation with sodium periodate, thus exposing free aldehyde residues on their surface, able to form Schiff bases with accessible hydrazide groups on the marker molecule. Fixed and oxidized erythrocytes could be stored for more than 1 year without losing their capacity to be agglutinated by polyhydrazides. The aggregating capacity of hydrazide-enriched molecules correlated well with their binding to oxidized cell surface moieties.

Aldehydes↗

Comparison of RNA hybridization, hemagglutination assay, titration of infectious virus and immunofluorescence as methods for monitoring influenza virus replication in vitro.

Rapid and sensitive methods for the monitoring of influenza virus replication in vitro are needed to address several research questions. Four methods based on different principles were compared: the hemagglutination (HA) assay, the measurement of virus infectivity titers in culture supernatants, the enumeration of infected cells by immunofluorescence and RNA hybridization techniques using digoxigenin (DIG) labeled RNA probes. To this end, MDCK cells were infected at different multiplicities of infection (moi) with a recent influenza A virus (A/Netherlands/18/94 H3N2) and the kinetics of virus replication were monitored with these four assays. At high moi, virus released into the culture supernatant of infected cells was detected by the HA assay 12 h post infection, whereas at lower moi (< or = 0.01) the first HA activity was not detected before 24 h post infection. The measurement of infectious viruses in the culture supernatant proved to be more sensitive, since 4-12 h post infection newly produced virus was detected depending on the moi used. This finding was in agreement with results obtained by the immunofluorescence assay using an antibody preparation specific for the nucleoprotein: single infected cells could be detected as early as 4 h post infection. At this time point, positive signals were also obtained when mRNA/cRNA specific hybridization was carried out for the NP gene segment, but not for viral NP RNA or RNA specific for the hemagglutinin, which were only detected at later time points after infection. Thus, besides direct measurement of infectious virus and immunofluorescence, RNA hybridization proved to be a sensitive assay for monitoring influenza virus replication in vitro.

Animals↗

An indirect hemagglutination antibody test to detect antibodies to Borrelia burgdorferi in patients with Lyme disease.

An indirect hemagglutination antibody (IHA) test was evaluated for its ability to detect borrelial antibodies in serum samples from patients with Lyme disease. The key test reagent developed for this antibody detection system was tannic acid-treated and glutaraldehyde-fixed sheep red blood cells (SRBC) containing Borrelia burgdorferi (Bb) antigens attached to the outer surface of the SRBC. In order to establish suitable cut-off titers, initial specificity and sensitivity measurements were made using sera from 100 anonymous healthy volunteers and 30 additional pre-determined samples known to be non-reactive or reactive for Lyme disease or syphilis. These results were compared with those obtained using a commercially available ELISA. At titers >/=64, the IHA test had a combined 98% specificity and 100% sensitivity for these 130 serum samples, 30 of which were known positives or negatives, whereas the ELISA was less specific (93%) and much less sensitive (80%). Subsequent testing was performed on sera from 65 patients with the erythema migrans (EM) rash and 20 patients with early disseminated (cardiac/neurologic) symptoms or with Lyme arthritis. At initial presentation, 46-48% of the EM patients had IHA reactivity, with titers >/=128, while 42% were positive in the ELISA. Follow-up testing performed on these EM patients, 8-12 days after receiving antibiotic treatment, revealed that Bb antibodies were detected best by the IHA test (83-86% reactive) relative to the ELISA (81% reactive). Bb antibodies were readily detectable on all of the serum samples from the early disseminated and late stage Lyme disease cases in both assay systems. Based on these results and because of its technical and interpretive simplicity, the IHA test should be considered as a useful and convenient alternative for the serological analysis of Bb infections.

Antibodies, Bacterial↗

Detection of antibodies to leptospiral genus-specific antigen in human and animal sera by indirect hemagglutination test with a partially purified genus-specific protein antigen.

Antibodies against leptospiral genus-specific antigen were detected in the sera from clinically diagnosed human leptospirosis and suspected animal leptospirosis by indirect hemagglutination (IHA) test with a partially purified genus-specific protein antigen (GP-Ag). The reaction was positive in the infected humans and animals irrespective of the leptospiral serovars. No significant correlation was found between IHA titer against GP-Ag and microscopic agglutination (MA) titer. IHA titer did not always develop in parallel with MA titers. Sera obtained from healthy individuals were negative in both IHA and MA tests.

Animals↗

The hemagglutination inhibition antibody responses to an inactivated influenza vaccine among healthy adults: with special reference to the prevaccination antibody and its interaction with age.

The immunogenicity of the trivalent split-virus influenza vaccine was investigated among 70 healthy adults (mean age: 48.5, range: 36-68). The vaccine antigens were: A/Yamagata/32/89 (H1N1); A/Beijing/352/89 (H3N2); and B/Bangkok/163/90. Regarding the entire sample, the vaccine induced a tenfold or more rise on the average in the hemagglutination inhibition (HAI) antibody to each antigen. The response rates (greater than or equal to a fourfold rise) were about 90% or more among those with a prevaccination titer < or = 1:64 (equivalent to < or = 1:16 on the Western scale; in Japan, the HAI titers are expressed by the final, and not the initial, dilution of the serum; from hereon our findings will be expressed using the Japanese scale), whereas they were 0-50% at > or = 1:128. Thus, the prevaccination titer was negatively associated with antibody induction. The achievement rates (postvaccination titer > or = 1:128) among those with a prevaccination titer < 1:16 remained at 48-68%. Regarding the analysis of variance, a significant effect on antibody induction was indicated for the prevaccination titer (P < or = 0.002), but not for age (P > or = 0.425). The interaction between the prevaccination titer and age was significant for A/Yamagata (P = 0.030), while it was also suggestive for A/Beijing (P = 0.054): as age increased, those with no preexisting antibody (< 1:16) showed greater titer rises, in contrast to the smaller rises among those with a titer > or = 1:16. Based on the attack survey conducted separately, the vaccine efficacy on influenza-like illnesses with fever < or = 37 degrees C and > or = 37.5 degrees C was calculated to be 16% (95% confidence interval: -66% to 57%) and 37% (-55% to 74%), respectively.

Adult↗

[Experimentation in Lille with the passive hemagglutination test for the detection of Australia antigen. Comparison with other methods].

Among 5347 blood donors sera, the reverse passive hemagglutination test (RPH) enabled to detect 3,55% of HB ag (whereas 2,43% by EID). On the other hand, among 180 patients, the rate of positive reactions was observed to be the same for both techniques, which must be attributed to the presence of higher titer antigens. In the two series all positive reactions found by RPH method were also tested by RIA; this enabled to eliminate a great number (18% and 27%) of false positive reactions, which remain the disadvantage of the method. It must be also noticed that the complement fixation reaction gave the same results as the RIA.

Blood Donors↗

Comparative evaluation of two hemagglutination tests for the detection of anti-Helicobacter pylori antibodies.

Two hemagglutination assays (IHAs) for detecting anti-H. pylori antibodies were evaluated. In 257 patients, both tests revealed sensitivity or specificity values between 71-79% or 81-82%, respectively, which were lower than those of an anti-H. pylori ELISA. Notably, the sensitivities of both IHAs varied in subgroups of patients between 48-95%. Based on the validation, the IgG-ELISA is superior to both IHAs tested, and therefore, should be used in routine diagnostics.

Adolescent↗

K-antigen identification, hemolysin production, and hemagglutination types of Escherichia coli O6 strains isolated from patients with urinary tract infections.

Serotyping of 1918 Escherichia coli strains isolated in significant cell numbers from the urine of patients with urinary tract infections (UTI) revealed the presence of 117 O6 strains. The K antigens were identified by means of K-specific phages and serological methods. The phages used included a K1 phage pool (phi 1, A-E) and the separate phages phi 2, phi 5, phi 7, phi 12 and phi 13. The presence of H antigens, type 1 fimbriae formation, hemolysin production and mannose-resistant hemagglutination (MRHA) ability with human A, sheep, calf and pig erythrocytes were also analyzed. Six different MRHA types were defined and discussed in relation to the O6:K:H serotype. Remarkably, E. coli O6 strains were found to possess a whole arsenal of virulence factors (K antigens, MRHA, hemolysin). The most common serotypes - O6:K2:H1/H- (26), O6:K5:H1/H- (35) and O6:K13:H1/H- (20) - differed from each other in some cases in both MRHA type and hemolysin production.

Antigens, Bacterial↗

Expression, purification, and characterization of an anti-RBCFab-p24 fusion protein for hemagglutination-based rapid detection of antibodies to HIV in whole blood.

A fusion protein A41Fabp246 consisting of Fab of an anti-RBC MAb A41 and amino acids 8-153 of HIV-1 p24 (p246) fused at the C-terminus of A41Fd was purified following assembly of A41Fdp246 with A41 LC in vitro, using a denaturation- renaturation protocol and a 4-step column chromatography procedure. The highly purified, monomeric A41Fabp246 was then evaluated for hemagglutination-based detection of anti-p24 antibodies using sera from HIV-infected individuals. This derivative of p24 is devoid of maximum homology region and the C-terminal domain of p24, which is responsible for oligomerization of p24, but retains full complement of immunodominant epitopes. This new fusion protein in combination with fusion proteins consisting of monovalent fragment of another anti-human RBC antibody fused to immunodominant regions of envelope glycoproteins of HIV-1 and HIV-2 should be useful in preparing a cocktail of reagents for highly sensitive detection of anti-HIV antibodies in whole blood.

Animals↗

Double aldehyde stabilisation of erythrocytes in the indirect hemagglutination for echinococcosis.

Sheep erythrocytes were stabilised with glutaraldehyde tanned and fixed with formalin in the indirect hemagglutination test (IHA-GF) and sensitised with hydatid antigen for the diagnosis of human cystic echinococcosis (CE). The sensitivity of this method was compared to that prepared with fresh tanned cells (IHA-TA) in 278 sera from hydatid patients. The sensitivity of IHA-GF (87.8%) was higher than that of IHA-TA (85.6%), the difference being insignificant. Higher geometric mean titres were obtained by IHA-GF (1:13300) than by IHA-TA (1:11600). The use of two sorts of aldehydes proved to be a satisfactory method, showing high sensitivity, a very good specificity and some advantages. The sensitised cells retained their diagnostic effectiveness for at least 15-18 months when stored at 4 degrees C. The technique is inexpensive and rapid, allowing the testing of a large number of sera. The method reduces the variation of the results and improves the reproducibility of the test. When the minimal diagnostic titre-1:400 is used the specificity of IHA-GF might increase by 2.9% while the sensitivity might decline by only 1.4%. The IHA-GF demonstrated better immunodiagnostic characteristics than enzyme linked immunosorbent assay (ELISA) and Latex agglutination test (LAT). The IHA-GF should be considered as an useful method in the range of classical diagnosis for the serology of CE. The clinical diagnostic potential should be increased by a combination of at least two tests: IHA-GF and ELISA or LAT.

Animals↗

A spin-label study on fusion of red blood cells induced by hemagglutinating virus of Japan.

Fusion of red blood cells (RBC) induced by hemagglutinating virus of Japan (HVJ) has been studied using a phosphatidylcholine spin label. The spin label was readily incorporated and diffused into the lipid bilayer portion of the viral envelope. The exchange broadening in the electron spin resonance (ESR) spectrum of densely labeled virus disappeared rapidly when the virus was mixed with RBC at 37 degrees. The spectrum gradually approached that of the host cell spin labeled with the phosphatidylcholine label. The results directly indicate transfer and intermixing of phospholipid molecules between the viral envelope and RBC membrane. The transfer reaction was strongly dependent on temperature. No transfer was observed at lower temperatures where the virus adsorbed to the cell and caused aggregation but no hemolysis and fusion. The transfer rate remained negligibly small until 19 degrees and increased rapidly between 25 and 30 degrees. The virus-induced hemolysis showed similar temperature dependence. The transfer rate was greatly reduced under inhibitory conditions of fusion: glutaraldehyde treatment of RBC, trypsin treatment of HVJ, or the presence of concanavalin A. Only slight transfer was observed from fusion-inactive influenza virus to RBC. The transfer was greatly enhanced by the help of HVJ. The close parallelism suggests that the transfer and intermixing are necessary steps to the cell fusion. The transfer rate was dependent on fluidity of the host cell membrane and independent of the viral dose. The virus-induced transfer of phospholipid molecules between RBC's was also detected by the spin label. Its temperature dependence was quite similar to that for the virus-to-cell transfer. The intercellular transfer was nearly proportional to the viral dose.

Cell Fusion↗